|
| Status |
Public on Jun 12, 2013 |
| Title |
51765Pre |
| Sample type |
RNA |
| |
|
| Source name |
skeletal muscle
|
| Organism |
Homo sapiens |
| Characteristics |
subject: 51765Pre
age: 18
time: prior to exercise training
tissue: skeletal muscle biopsy
|
| Treatment protocol |
Subjects were physiologically assessed at baseline
|
| Growth protocol |
Not applicable
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Paired tissue samples (obtained before and after exercise training) of ~20 mg each were processed simultaneously in 1 mL TRIzol (Invitrogen) using a Mini-Beadbeater-8 (Biospec Inc.) for 15 sec on the homogenize setting. After 5 min of incubation at room temperature, 200 µL of chloroform (Sigma-Aldrich) was added and samples shaken vigorously by hand. Samples were briefly incubated on ice prior to centrifugation at 12,000 g for 15 min. The supernatant was removed and mixed with isopropanol (Sigma-Aldrich) and spun once more at 12,000 g for 10 min, after 10 min of incubation. After a single washing step with 75% EtOH RNA pellets were re-suspended in 40 µL DEPC-treated water (Ambion) and quantified using a NanoDrop Spectrophotometer (NanoDrop Technologies).
|
| Label |
biotin
|
| Label protocol |
Reverse transcription of RNA was carried out using the Affymetrix 3′ IVT express kit. 100 ng of total RNA was reverse transcribed as per manufacturer's protocol, and quantified using a Nanodrop ND-1000 instrument. aRNA was fragmented and labeled as per manufacturers protocol and hybridized to Affymetrix U133+2 arrays (Affymetrix, USA).
|
| |
|
| Hybridization protocol |
Arrays were washed, stained and scanned following Affymetrix standard procedures, using an Affymetrix 3000 7G scanner and Affymetrix 450 wash station. A visual inspection of each array was carried out.
|
| Scan protocol |
Affymetrix Gene ChIP Scanner 3000 7G
|
| Description |
gene expression prior to exercise training
HER_95_51765Pre
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling as normalization method.
|
| |
|
| Submission date |
Jun 12, 2013 |
| Last update date |
May 30, 2024 |
| Contact name |
James Timmons |
| E-mail(s) |
jamie@augurprecisionmedicine.com, jamie.timmons@gmail.com
|
| Organization name |
Augur Precision Medicine LTD
|
| Street address |
Bridge of Allan
|
| City |
Stirling |
| ZIP/Postal code |
FK9 4AX |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE47874 |
The Heritage (HEalth, RIsk factors, exercise Training And GEnetics) family study |
|
